Production of a recombinant membrane protein in an Escherichia coli strain for the whole cell biosynthesis of phenylacetic acids

Oelschlägel, Michel; Heiland, Claudia; Schlömann, Michael; Tischler, Dirk

doi:10.1016/j.btre.2015.05.002

Cited by 18 publications

(17 citation statements)

References 38 publications

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“…Nevertheless, small amounts of 2-phenylethanol have been reported using E. coli cells as whole cell biocatalyst ( Panke et al, 2000 ; Panke et al, 2002 ). This ethanol formation is contributed to unspecific reactions within the cells and not (or only in a low extent) caused directly by the SMO itself because 2-phenylethanol formation has also been reported by another study using a recombinantly expressed SOI in E. coli in presence of styrene oxide without an SMO ( Oelschlägel et al, 2015a ). No by-product formation was proven using the styC knock-out strain of Pseudomonas sp.…”

Section: More Details On the Enzymes Of Side-chain Oxygenation And Thsupporting

confidence: 64%

“…This was also initiated in order to produce phenylacetic acid from styrene without – in contrast to the application of native styrene degraders – further product metabolization. Initial studies investigated the transformation of styrene oxide by an SOI in E. coli BL21(DE3) in order to produce phenylacetaldehyde from styrene oxide ( Oelschlägel et al, 2015a ). Because E. coli harbors an own PAD (FeaB), the modification of this E. coli strain by introducing a single SOI gene already lead to the formation of phenylacetic acid as main product ( Oelschlägel et al, 2015a ).…”

Section: The Biotechnological Potential Of the Side-chain Oxygenationmentioning

confidence: 99%

“…Initial studies investigated the transformation of styrene oxide by an SOI in E. coli BL21(DE3) in order to produce phenylacetaldehyde from styrene oxide ( Oelschlägel et al, 2015a ). Because E. coli harbors an own PAD (FeaB), the modification of this E. coli strain by introducing a single SOI gene already lead to the formation of phenylacetic acid as main product ( Oelschlägel et al, 2015a ). Phenylacetaldehyde occurs only as an intermediate and undergoes further oxidation by FeaB during these experiments.…”

Section: The Biotechnological Potential Of the Side-chain Oxygenationmentioning

confidence: 99%

“…Phenylacetaldehyde occurs only as an intermediate and undergoes further oxidation by FeaB during these experiments. A further gene-knock-out has been discussed to enhance the yield of phenylacetaldehyde ( Oelschlägel et al, 2015a ). Nevertheless, phenylacetaldehyde represents also a toxic compound for the E. coli cells at concentrations of 1.35 mM which limits this way of application ( Oelschlägel et al, 2015a ).…”

Section: The Biotechnological Potential Of the Side-chain Oxygenationmentioning

confidence: 99%

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A Review: The Styrene Metabolizing Cascade of Side-Chain Oxygenation as Biotechnological Basis to Gain Various Valuable Compounds

2018

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Styrene is one of the most produced and processed chemicals worldwide and is released into the environment during widespread processing. But, it is also produced from plants and microorganisms. The natural occurrence of styrene led to several microbiological strategies to form and also to degrade styrene. One pathway designated as side-chain oxygenation has been reported as a specific route for the styrene degradation among microorganisms. It comprises the following enzymes: styrene monooxygenase (SMO; NADH-consuming and FAD-dependent, two-component system), styrene oxide isomerase (SOI; cofactor independent, membrane-bound protein) and phenylacetaldehyde dehydrogenase (PAD; NAD+-consuming) and allows an intrinsic cofactor regeneration. This specific way harbors a high potential for biotechnological use. Based on the enzymatic steps involved in this degradation route, important reactions can be realized from a large number of substrates which gain access to different interesting precursors for further applications. Furthermore, stereochemical transformations are possible, offering chiral products at high enantiomeric excess. This review provides an actual view on the microbiological styrene degradation followed by a detailed discussion on the enzymes of the side-chain oxygenation. Furthermore, the potential of the single enzyme reactions as well as the respective multi-step syntheses using the complete enzyme cascade are discussed in order to gain styrene oxides, phenylacetaldehydes, or phenylacetic acids (e.g., ibuprofen). Altered routes combining these putative biocatalysts with other enzymes are additionally described. Thus, the substrates spectrum can be enhanced and additional products as phenylethanols or phenylethylamines are reachable. Finally, additional enzymes with similar activities toward styrene and its metabolic intermediates are shown in order to modify the cascade described above or to use these enzyme independently for biotechnological application.

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Section: More Details On the Enzymes Of Side-chain Oxygenation And Thsupporting

confidence: 64%

Section: The Biotechnological Potential Of the Side-chain Oxygenationmentioning

confidence: 99%

Section: The Biotechnological Potential Of the Side-chain Oxygenationmentioning

confidence: 99%

Section: The Biotechnological Potential Of the Side-chain Oxygenationmentioning

confidence: 99%

See 2 more Smart Citations

A Review: The Styrene Metabolizing Cascade of Side-Chain Oxygenation as Biotechnological Basis to Gain Various Valuable Compounds

2018

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“…strain ADP1 to gain high yields of soluble protein during expression in Escherichia coli or, if necessary, in Acinetobacter sp. ( Oelschlägel et al, 2015 ). After ligation of the genes into the vector pET16bP ( Table 1 ), the resulting recombinant plasmids pS Ro OYE2_P01 and pS Ro OYE2a_P01 were transformed into E. coli BL21 (DE3).…”

Section: Methodsmentioning

confidence: 99%

Functional characterization and stability improvement of a ‘thermophilic-like’ ene-reductase from Rhodococcus opacus 1CP

et al. 2015

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Ene-reductases (ERs) are widely applied for the asymmetric synthesis of relevant industrial chemicals. A novel ER OYERo2 was found within a set of 14 putative old yellow enzymes (OYEs) obtained by genome mining of the actinobacterium Rhodococcus opacus 1CP. Multiple sequence alignment suggested that the enzyme belongs to the group of ‘thermophilic-like’ OYEs. OYERo2 was produced in Escherichia coli and biochemically characterized. The enzyme is strongly NADPH dependent and uses non-covalently bound FMNH2 for the reduction of activated α,β-unsaturated alkenes. In the active form OYERo2 is a dimer. Optimal catalysis occurs at pH 7.3 and 37°C. OYERo2 showed highest specific activities (45-50 U mg-1) on maleimides, which are efficiently converted to the corresponding succinimides. The OYERo2-mediated reduction of prochiral alkenes afforded the (R)-products with excellent optical purity (ee > 99%). OYERo2 is not as thermo-resistant as related OYEs. Introduction of a characteristic intermolecular salt bridge by site-specific mutagenesis raised the half-life of enzyme inactivation at 32°C from 28 to 87 min and improved the tolerance toward organic co-solvents. The suitability of OYERo2 for application in industrial biocatalysis is discussed.

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Biotechnological Applications of Styrene-Degrading Microorganisms or Involved Enzymes

Tischler

2015

SpringerBriefs in Microbiology

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Production of a recombinant membrane protein in an Escherichia coli strain for the whole cell biosynthesis of phenylacetic acids

Abstract: Graphical abstract

Cited by 18 publications

References 38 publications

A Review: The Styrene Metabolizing Cascade of Side-Chain Oxygenation as Biotechnological Basis to Gain Various Valuable Compounds

A Review: The Styrene Metabolizing Cascade of Side-Chain Oxygenation as Biotechnological Basis to Gain Various Valuable Compounds

Functional characterization and stability improvement of a ‘thermophilic-like’ ene-reductase from Rhodococcus opacus 1CP

Biotechnological Applications of Styrene-Degrading Microorganisms or Involved Enzymes

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